Development of chromosome sorting techniques using a dual laser chromosome sorter

我们已经开发了染色体分选技术，使用双激光染色体分选器来分析人类染色体的结构。用Hoechst 33258 （Ho）和chromomycin A3 （CA3）染色的人类染色体依次流过两束激光，一束调节到457.9nm激发CA3，另一束调节到351.1-363.8nm激发Ho。单个染色体的Ho和CA3含量是通过测量它们通过两束激光发出的荧光强度来确定的。这些测量值累积起来形成荧光强度分布。分选技术已经从能够用单个激光系统分选10-20个人类染色体片段发展到能够用双激光系统分选21个片段，包括15种独特的类型。d此外，从人类细胞中分离的染色体不能以高纯度分类的染色体类型可以从含有一条或几条人类染色体的人-仓鼠杂交细胞系中分类。

We have developed chromosome sorting techniques using a dual laser chromosome sorter to analyze the structure of human chromosomes.Human chromosomes stained with Hoechst 33258 (Ho) and chromomycin A3 (CA3) flowed sequentially through two laser beams, one adjusted to 457.9nm to excite CA3 and the other adjusted to 351.1-363.8nm to excite Ho. The Ho and CA3 contents of individual chromosomes were determined by measuring the intensities of fluorescence that were emitted as they passed two laser beams. These measurements were accumulated to form fluorescence intensity distributions.Sorting techniques have evolved from the ability to sort 10-20 human chromosome fractions with a single laser system to sorting 21 fractions including 15 unique types with a dual laser system.dFurthermore, chromosome types that cannot be sorted at high purity from chromosomes isolated from human cells can be sorted from human-hamster hybrid cell lines containing one or a few human chromosomes.